Accumulation and localization of troponin-T in developing hearts of Ambystoma mexicanum

Development ◽  
1984 ◽  
Vol 84 (1) ◽  
pp. 1-17
Author(s):  
Rebecca A. Fuldner ◽  
Soo-Siang Lim ◽  
Marion L. Greaser ◽  
Larry F. Lemanski
Keyword(s):  
Monoclonal Antibodies ◽  
Indirect Immunofluorescence ◽  
Gel Electrophoresis ◽  
Troponin T ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Composition ◽  
Ambystoma Mexicanum ◽  
Striking Difference ◽  
Lethal Mutant ◽  
Mutant Cells

Troponin-T (Tn-T) expression in developing hearts of axolotls, Ambystoma mexicanum, was studied with the use of polyclonal and monoclonal antibodies and SDS-polyacrylamide gel electrophoresis. In precontractile hearts (stage 32/33), Tn-T was present in addition to myosin, actin and tropomyosin as evidenced by the presence of the protein bands in SDSgels and by indirect immunofluorescence. Tn-T was localized in amorphous collections at the peripheries of these precontractile cells. Hearts of normal and cardiac lethal mutant siblings were also analysed for Tn-T expression. No detectable differences in the quantity of protein present was observed by gel electrophoresis or by indirect immuno-fluorescence. The most striking difference concerned the localization of the protein. In normal hearts, Tn-T was primarily localized in the I-bands of organized myofibrils; however, in mutant cells the Tn-T was localized in amorphous collections at the cell peripheries suggesting a reduction of myofibrillar organization in these cells. No differences were observed in the contractile protein composition between normal and mutant em-bryonic hearts by gel electrophoresis experiments.

Monoclonal antibodies against surface antigens of schistosomula of Schistosoma mansoni

Parasitology ◽  
1982 ◽  
Vol 84 (1) ◽  
pp. 65-82 ◽  
Author(s):  
D. W. Taylor ◽  
A. F. Butterworth
Keyword(s):  
Monoclonal Antibodies ◽  
Gel Electrophoresis ◽  
Primary Infection ◽  
Polyacrylamide Gel Electrophoresis ◽  
Apparent Molecular Weight ◽  
Surface Antigens ◽  
Soft Agar ◽  
Surface Proteins ◽  
Spleen Cells

SUMMARYMonoclonal antibodies have been produced after fusion of NS-1 murine myeloma cells with spleen cells from mice immunized either by chronic primary infection or with irradiated cercariae: in both cases, animals were challenged with live cercariae 7 days before fusion. The initial cultures were screened for anti-schistosomular antibodies both by a radioimmunoassay with whole schistosomulum extracts and by immunofluorescence. There was no correlation between the two techniques and subsequent screening was carried out by immunofluorescence. Cloning was carried out in soft agar and 7 cloned cell lines, from 5 initial cultures, were selected for detailed study. Products of 6 of these 7 lines were monoclonal, as judged by isoelectricfocusing of [35S]methionine-labelled supernatant fluids, and their binding to live schistosomula was specific. None of the antibodies showed detectable activity in mediating eosinophil- or complement-dependent damage to schistosomula in vitro. However, 2 antibodies were successfully used to isolate surface proteins with an apparent molecular weight of 24000 on SDS-polyacrylamide gel electrophoresis.

Protein composition of Methanococcus thermolithotrophicus flagella

1992 ◽  
Vol 38 (11) ◽  
pp. 1162-1166 ◽  
Author(s):  
Alla S. Kostyukova ◽  
Georgi M. Gongadze ◽  
Anna Ya. Obraztsova ◽  
Konstantin S. Laurinavichus ◽  
Oleg V. Fedorov
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Key Words ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Composition ◽  
Complete Removal ◽  
Methanococcus Thermolithotrophicus ◽  
Molecular Weights ◽  
Dodecyl Sulfate

Sodium dodecyl sulfate – polyacrylamide gel electrophoresis of flagella from the thermophilic methanogen Methanococcus thermolithotrophicus indicated that they were composed of three major proteins, with molecular weights of 62 000,44 000, and 26 000, whereas all previously studied flagella of mesophilic methanogens consisted of two subunits. Proteins were isolated by preparative electrophoresis followed by complete removal of sodium dodecyl sulfate and their renaturation. It was shown that at least two of the proteins contain a thermostable domain whose complete denaturation proceeds only upon prolonged boiling in the presence of sodium dodecyl sulfate. Key words: flagellin, thermostability, archaebacteria, Methanococcus thermolithotrophicus.

Analysis of proteins in rabbit pulmonary surfactant using monoclonal antibodies

1986 ◽  
Vol 250 (3) ◽  
pp. C460-C467 ◽  
Author(s):  
R. J. King ◽  
H. M. Martin ◽  
J. B. Baseman ◽  
J. Morrison-Plummer
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Pulmonary Surfactant ◽  
Polyacrylamide Gel Electrophoresis ◽  
Low Molecular Weight ◽  
Weight Group ◽  
Molecular Weights ◽  
Intermediate Group

We have used monoclonal antibodies developed against the apolipoproteins associated with pulmonary surfactant purified from rabbit lavage fluid to study the expression of epitopes common to these proteins. The pulmonary surfactant contained nearly 20 proteins, of which at least 10 were not derived from serum. Electrophoresis, with sulfhydryl reduction of these proteins indicated apparent molecular weights of approximately 155, 135, 125, and 115 X 10(3) (high-molecular-weight group); 80, 70, and 60 X 10(3) (intermediate group); and 18 through 10 X 10(3) (low-molecular-weight group). Two-dimensional polyacrylamide gel electrophoresis, in which the proteins were electrophoresed without reduction in the first dimension, but with sulfhydryl reduction in the second dimension, revealed that the 80, 70, and 60 X 10(3) proteins dissociated into proteins of nominal molecular weights of 40, 35, and 30 X 10(3), respectively. In contrast, the 125 and 115 X 10(3) proteins of the high-molecular-weight group contained a protein which could only be reduced to a minimum molecular weight of 55 to 60 X 10(3). Monoclonal antibodies generally were of three types: those that reacted strongly with the high-molecular-weight group and weakly with the intermediate group; those that reacted conversely; and those that reacted only with the low-molecular-weight group. Our results indicate that at least two different surfactant apolipoproteins, with differing minimum molecular weights in SDS-polyacrylamide gel electrophoresis, have common epitopes. Although these results cannot certify a physiological relationship between these proteins, they suggest that the intracellular synthesis or extracellular processing of surfactant apolipoproteins may be more complicated than predicted by the findings of previous experiments, perhaps involving the posttranslational assembly of one surfactant protein into oligomers which resist dissociation under the conditions used for the analyses.

Yolk proteins in salmon (Salmo salar) oocytes, eyed eggs, and alevins differing in viability

1990 ◽  
Vol 68 (5) ◽  
pp. 895-900 ◽  
Author(s):  
Thomas Olin ◽  
Alexandra von der Decken
Keyword(s):  
Molecular Weight ◽  
Salmo Salar ◽  
Gel Electrophoresis ◽  
Developmental Stages ◽  
Polyacrylamide Gel Electrophoresis ◽  
Specific Antigen ◽  
Protein Composition ◽  
Polyacrylamide Gel ◽  
Parental Origin ◽  
Yolk Proteins

The developmental stages of oocytes, eyed eggs, and alevins from salmon (Salmo salar) were compared for their yolk protein composition. In oocytes, SDS–polyacrylamide gel electrophoresis showed high amounts of a protein with the molecular weight (Mr) of 94 000. In eyed eggs, the 94 000 protein decreased and was undetectable in the alevins. Furthermore, in eyed eggs the proteins of 67 000, 30 000, and 27 000 increased, while in the alevins the concentration of the 67 000 protein decreased and that of the 39 000 increased. Vitellogenin-specific antigen sites analyzed by immunoblotting were most pronounced with the proteins of 94 000, 67 000, 39 000, 30 000, 23 000, and 19 000. Separation of the yolk proteins by HPLC gave four peaks at 280 nm for all three developmental stages. Each peak consisted of several proteins as analyzed by SDS–polyacrylamide gel electrophoresis. The 7-day-old alevins sampled from groups of different parental origin showed differences in the amount of the 67 000 and 23 000 proteins. Expectancy of survival within the group in connection with a slow disappearance of the 67 000 and 23 000 proteins was statistically significant. A fast disappearance may be used as an indication of, but not as the reason for, a high mortality within one group of alevins.

Evaluation of wheat endosperm protein fingerprints as indices of Udon-noodle making quality

2001 ◽  
Vol 52 (9) ◽  
pp. 919
Author(s):  
H. Nakamura
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Composition ◽  
Protein Band ◽  
Eating Quality ◽  
Endosperm Protein ◽  
Wheat Lines ◽  
Noodle Quality

Hexaploid wheat (Triticum aestivum) endosperm protein fingerprints were used to determine the indices of Japanese soft Udon-noodle quality. The endosperm proteins of Japanese Udon wheat lines were fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis to determine differences between them in protein composition in two soil environments. The differences between the lines included differences in the composition of the endosperm with regard to the 53 kDa protein band or high-molecular-weight glutenin subunit (HMW-GS) 2* and in the sensory viscoelasticity score of cooked noodles, which is related to eating-quality in Japanese Udon wheats. One line (Kanto 107) showed variation for the presence of the 53 kDa and HMW-GS 2* bands between environments.

Comparison of the cell envelope proteins of Micrococcus cryophilus with those of Neisseria and Branhamella species

1976 ◽  
Vol 22 (2) ◽  
pp. 309-312 ◽  
Author(s):  
R. R. B. Russell ◽  
I. J. McDonald
Keyword(s):  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Gel Electrophoresis ◽  
Cell Envelope ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Composition ◽  
Polyacrylamide Gel ◽  
Outer Membranes ◽  
Branhamella Catarrhalis

In an attempt to elucidate the relation between Micrococcus cryophilus, Neisseria caviae, Neisseria ovis, and Branhamella catarrhalis, fractions derived from outer membranes of a strain of each organism were examined for protein composition by SDS – polyacrylamide gel electrophoresis. Micrococcus cryophilus outer membrane protein showed extensive similarities to that of N. ovis and contained a heat-modifiable protein which behaved almost identically with the corresponding bands previously shown to exist in N. caviae and N. oris. Branhamella catarrhalis protein was distinctly different from those of M. cryophilus and the two 'false neisserias' N. caviae and N. oris.

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